Expression of prostate specific antigen on the surface of a filamentous phage
| Autor: | Hans Lilja, Petri Saviranta, M. Karp, Kim Pettersson, R. Eerola, T Lövgren |
|---|---|
| Rok vydání: | 1994 |
| Předmět: |
Inovirus
Phage display medicine.drug_class viruses Phagemid Genetic Vectors Molecular Sequence Data Biophysics urologic and male genital diseases Monoclonal antibody Biochemistry Epitope Bacteriophage Antigen medicine Humans Trypsin Cloning Molecular Molecular Biology DNA Primers biology Base Sequence Antibodies Monoclonal Cell Biology Prostate-Specific Antigen biology.organism_classification Molecular biology Biotinylation |
| Zdroj: | Biochemical and biophysical research communications. 200(3) |
| ISSN: | 0006-291X |
| Popis: | We have constructed two phagemid vectors containing the gene coding for human Prostate Specific Antigen (PSA) translationally fused to a pel B signal sequence and minor coat protein III of phage fd leading to phage particles that carry PSA on their tips. Phages were characterized by Western blotting and by panning, using biotinylated monoclonal anti-PSA antibodies immobilized onto streptavidin-coated microtitration wells. Binding of PSA-phages was 10 2 - to 5×10 3 -fold more effective than that of wildtype phages. Trypsin treatment of the phage abolished the specific binding. Competitive panning with different concentrations of added purified PSA demonstrated that binding was PSA specific. Finally, Western-blot analysis confirmed that full-length and shorter degradation products of the fusion between PSA and protein III were visible as probed with anti-PSA antibody. |
| Databáze: | OpenAIRE |
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